CN104560760A - Pathogenic bacterium sorting and picking method in blood examination - Google Patents

Abstract

A pathogenic bacterium sorting and picking method in blood examination is characterized in that suspected pathogenic microbes in blood are inoculated and cultured many times, and then are sorted, the microbes are distinguished by using the growth characteristics of different varieties of microbes after enlarged culture and the characteristics of bacterium colonies, and suspected pathogenic bacteria are picked and undergo culture separation, so the effect of competitors is greatly eliminated, and the blood testing accuracy is improved. The method has the advantages of low cost, easy operation and implementation and good repeatability, and can overcome the limitations of traditional clinical examination to a certain extent.

Description

A kind of method of pathogenic bacterium sorting picking in blood test

Technical field

The present invention relates to medical science disease microorganism authentication technique field, specifically provide simply a kind of and the method for pathogenic bacterium sorting picking in blood test accurately.

Background technology

Blood is in the recycle system, the one tissue that heart and Endovascular circulate.Blood tissues is the one of reticular tissue.Be made up of blood plasma and hemocyte.Blood plasma includes various nutritive ingredient and inorganic salt, oxygen, hormone, enzyme, antibody and the products of cellular metabolism etc. such as plasma proteins (albumin, sphaeroprotein, Fibrinogen), lipoprotein.Hemocyte has red blood corpuscle, white cell and thrombocyte.Mammiferous blood has clotting mechanism, during plaque rupture, can by originally can be water-soluble in blood plasma fibrinogen and hemocyte etc. solidify out into blood cake, remaining transparent liquid is just called serum.

The physiological change of organism and pathological change often cause the change of blood ingredient, so the detection of blood ingredient has important clinical meaning.The blood of human normal be do not have germy.After human body is subject to bacteriological infection, when resistibility reduces, bacteriological infection human body generally first causes the inflammation of certain part, as furuncle, carbuncle, pneumonia, encephalitis etc.Medically the illness entering circulation of blood by local inflammation position because of bacterium is called microbemia.At this moment often have a small amount of bacterium and invade blood, if the systemic conditions of people is fine, this small amount of bacterium can be eliminated by the white corpuscle in the various antibody, phagocytic cell and the blood that produce in health.And if angioinvasive germ is a lot, toxicity is very strong, or due to human body overwork, malnutritive, severe trauma or have the reasons such as other diseases and resistibility is extremely low time, bacterium also can produce hazardous and noxious substances, with flow propagation to whole body by amount reproduction.At this moment patients is the serious toxicity symptom such as high heat, shiver with cold, headache, body temperature Chang Gaoda more than 40 DEG C.Meanwhile, bacterium can invade inflammation, the suppuration that each internal organs cause internal organs.If send out skin, various fash can be there is.The illness that after medically bacterium being entered blood, growth and breeding causes in blood calls septicemia.If after suppurative bacterium enters blood, not only breed in blood, cause some internal organs to form the bad or ypotension of abscess or even organ dysfunction, perfused tissue, this illness is then referred to as pyemia.Sepsis case fatality rate is higher, and septic shock and many organ pipes functional defect can occur the sepsis patient of nearly 9%, and death over half in intensive care unit is caused by septic shock and multiple organ dysfunction.Research shows, occur internal organs organ failure, shock, multiple infection, serious potential disease patient's prognosis poor.A common ground of these three kinds of illnesss of above microbemia, septicemia, pyemia has bacterium in blood samples of patients.If cultivated by blood samples of patients, namely pathogenic bacterium can be detected.Therefore, the Clinics and Practices of result to disease of blood microbial culture and qualification has very important clinical effect.

Hemoculture inspection detects microbemia and a kind of simple, the most the most frequently used method of fungemia, is the etiology basis confirming body bloodstream infection.The method of current general blood microbial culture is that the sample process gathered by medical test person is hatched, cultivated, and tentatively judges, and then sends report according to the Identification of Biological Characteristics of various pathogenic bacteria.But in actual clinical operation, the blood sample of collection directly hatches cultivation, usually there will be that some specific pathogenic bacterium content is very few, purity is too low and fail situation about detecting.Especially in recent years, the good hospital of many medical conditions adopts the equipment of automatization to carry out blood microbial culture substantially, the positive rate that there is hemoculture is at large not high enough, culture cycle is long, easily occur that pollution leads to errors the problems such as result, this all has very tremendous influence to selecting medication, timely and effectively infection control and last patient effect in clinical treatment.

Therefore provide a kind of simple accurately and the method that can improve pathogenic bacterium purity in blood test has extremely important meaning.

Summary of the invention

Produce for existing the technical problem existed, the object of this invention is to provide a kind of method of pathogenic bacterium sorting picking in blood test simple to operate, safe and reliable, with low cost.

The inventive method adopts traditional microbiological method for culturing and separating and hematology testing method to combine, thus reaches by target pathogenic bacterium sorting picking out.Concrete technical scheme is the method for bacterium sorting picking in a kind of blood test, it is characterized in that comprising step:

1) sampling inoculation: get blood sample of patient as sample using aseptic technique, streak inoculation, on substratum, is inverted in constant temperature culture 1 ~ 2d in 37 DEG C of constant incubators, observes colony growth situation;

2) sorting: by single for the doubtful bacterium after inoculation culture colony inoculation on serum-based basal culture medium flat board, 37 DEG C of constant temperature culture 1 ~ 2d, observe colony growth situation;

3) get bacterium: picking disease bacterium, identify further to be ready for use on other.

The present invention is by repeatedly carrying out sorting after inoculation culture to pathogenic microorganism doubtful in blood, the growth characteristics that after utilizing enlarged culturing, different types of microorganism is different and colony characteristics make it be distinguished and the doubtful bacterium of picking disease carries out culture of isolated, eliminate the impact of miscellaneous bacteria largely, improve the accuracy of blood sample test.The inventive method has enforcement with low cost, easy to operate, the advantage such as reproducible, can overcome the limitation of traditional clinical inspection to a certain extent.

Embodiment

Below with reference to specific embodiment, the present invention is described in further details.

Embodiment

Test raw material and reagent

1) blood sample of patient;

2) serum-based basal culture medium: serum homogeneous heating to 56 DEG C is kept 30min, and be stored in 4 DEG C, mix afterwards with minimum medium, minimum medium accounts for 80% ~ 90%, and serum accounts for 10% ~ 20%.Dual anti-stock solution (penicillin+Streptomycin sulphate) is added by 1% volume fraction;

Testing sequence:

1) sampling inoculation: get blood sample of patient as sample using aseptic technique, streak inoculation, on substratum, is inverted in constant temperature culture 1 ~ 2d in 37 DEG C of constant incubators, observes colony growth situation;

2) sorting: by single for the doubtful bacterium after inoculation culture colony inoculation on serum-based basal culture medium flat board, 37 DEG C of constant temperature culture 1 ~ 2d, observe colony growth situation;

3) get bacterium: picking disease bacterium, identify further to be ready for use on other.

Claims (1)

1. the method for pathogenic bacterium sorting picking in blood test, is characterized in that comprising step:

1) sampling inoculation: get blood sample of patient as sample using aseptic technique, streak inoculation, on substratum, is inverted in constant temperature culture 1 ~ 2d in 37 DEG C of constant incubators, observes colony growth situation;

2) sorting: by single for the doubtful bacterium after inoculation culture colony inoculation on serum-based basal culture medium flat board, 37 DEG C of constant temperature culture 1 ~ 2d, observe colony growth situation;

3) get bacterium: picking disease bacterium, identify further to be ready for use on other.